清开灵注射液诱发类过敏反应作用及其机制研究

Effect of Pseudoallergic Reactions of Qingkailing Injections and Its Mechanisms

关键词:  清开灵注射液  类过敏反应  机制

Key words:  qingkailing injection  pseudoallergic reactions  mechanisms

摘要:

目的 研究清开灵注射液诱发类过敏反应的作用及其机制。方法 采用BALB/c小鼠和BN大鼠，观察给予清开灵注射液后小鼠皮肤血管渗透性以及大鼠平均动脉血压等类过敏指标的变化；利用RBL-2H3大鼠嗜碱性粒细胞白血病细胞，观察清开灵注射液作用后磷脂酰肌醇3-激酶（PI3K）活性以及Rac1、PAK1、LIMK1和人肌动蛋白素（Cofilin）蛋白含量的变化。结果 5 mL·kg^-1清开灵注射液即可引起小鼠以血管通透性增高为主要表现的较为轻微的类过敏反应，而7.5 mL·kg^-1和15 mL·kg^-1清开灵注射液均可引起较为严重的类过敏反应，其反应呈剂量依赖性；另外，5，7.5和15 mL·kg^-1清开灵注射液可在一定时间范围内剂量依赖性地引起大鼠平均动脉血压降低。清开灵注射液体外可诱导RBL-2H3细胞发生类过敏反应，PI3K激酶活性增加，Rac1、PAK1、LIMK1和Cofilin蛋白含量增高，且具有剂量依赖性。结论 小鼠和大鼠体内类过敏反应检查法和RBL-2H3细胞中PI3K激酶及Rac1、PAK1、LIMK1、Cofilin蛋白含量检查法可作为类过敏反应检查方法。初步认定清开灵注射液诱导类过敏反应的发生可能是基于PI3K-Rac1细胞运动信号转导通路。

Abstract:

OBJECTIVE To study the effect of pseudoallergic reactions of Qingkailing injections and its mechanisms. METHODS The BALB/c mice and BN rats were used to observe the changes of vascular permeability, mean arterial blood pressure(MABP) and other pseudoallergic reactions indicators with Qingkailing injection. RBL-2H3 cells were stimulated with Qingkailing injections and then were used to observe the activity of PI3K and the protein expression of Rac1, Pak1, Limk1 and cofilin. RESULTS The 5 mL·kg^-1 Qingkailing injection could cause to vascular permeability increase in mice as the main performance of the pseudoallergic reactions, while 7.5 mL·kg^-1 and 15 mL·kg^-1 Qingkailing injection caused serious pseudoallergic reactions in mice. Qingkailing injection in 5, 7.5 and 15 mL·kg^-1 caused rats' MABP to reduce in a dose dependent manner. In vitro, Qingkailing injection can induce pseudoallergic reactions and increase in levels of PI3K, Rac1, PAK1, LIMK1 and cofilin in a dose dependent manner in RBL-2H3 cells. CONCLUSION The tests in mice, rats and the content test of PI3K kinase, Rac1, PAK1, LIMK1 and cofilin in RBL-2H3 cells can be used as kinds of pseudoallergic reactions tests. Pseudoallergic reactions induced by Qingkailing injection is likely to be based on PI3K-Rac1 signal transduction pathways.